


Vol 472, No 1 (2017)
- Year: 2017
- Articles: 22
- URL: https://journal-vniispk.ru/1607-6729/issue/view/13161
Biochemistry, Biophysics, and Molecular Biology
Transcriptomic changes in human umbilical cord blood endothelial cells under simulated microgravity
Abstract
Microarray analysis of cultured endothelial cells was performed 24 h after simulated microgravity. A significant change in the expression of 177 genes that can be classified into several functional clusters was detected. Among the genes with overexpression, clusters of cell response to external stimuli and regulation of cell motility and proliferation can be reliably distinguished. Among down-regulated genes, clusters of transcription factors with the “zinc fingers” domain and factors involved in the regulation of morphogenesis and angiogenesis were distinguished. The overlapping of signaling pathways involved in mechanotransduction and inflammatory changes is discussed.



The level of proapoptotic gene transcripts in wheat leaves under high temperature stress
Abstract
Under exposure of wheat plants to high temperatures (33, 37, and 43°C), the level of transcripts of the genes encoding proapoptotic proteins—TaMCAII (encodes type II metacaspase) and TaBAX (functional homologue of the CDF1 gene of Arabidopsis thaliana)—in leaf cells increased. This process at temperatures of 37 and 43°C passed ahead of (in the case of TaBAX mRNA) or was accompanied by (in the case of TaMCAII mRNA) the emergence of signs of nucleosomal DNA degradation in leaves. The accumulation of malondialdehyde in the leaves of seedlings exposed to temperatures of 37 and 43°C coincided in time with a change in the TaMCAII gene expression and the emergence of signs of programmed cell death.



Flow cytometric analysis of the ERCC1, marker of DNA damage repair, in the tumor specimens embedded into paraffin blocks
Abstract
The differences in expression of ERCC1 were estimated between tumor specimens embedded into paraffin blocks and surgical biopsy specimens of non-small cell lung cancer as well as breast and ovarian cancers. Concordance or differences not higher than 20% were observed in 73% of the cases. The number of the cases with more significant differences in ERCC1 expression was less than 17%. The results show that ERCC1 detection in surgical biopsy specimens by flow cytometry is the more preferable method due to reduced preanalytical phase of the analysis.



Regeneration of jejunal wall defect using an implant based on silk fibroin fibers
Abstract
Regenerative properties of fibroin implant vitalized with allogeneic bone marrow cells were assessed. The study was performed using the experimental model of rat jejunum wall damage. Three weeks after surgery, we observed recovery of all layers of the jejunum wall at the site of injury and complete degradation of the implant material.



Genomic mapping of chromatin proteins by using Daminv modification of an FLP-dependent DamID approach
Abstract
To identify interactions of chromatin proteins with the genome of the cell type of interest that is a part of heterologous tissues and organs of Drosophila, an FLP-dependent DamID approach was recently developed [4], which does not require sorting of cells or nuclei. Here, a modification of this approach, Daminv, is described. The modified approach was validated by generating the binding pattern of the LAM protein, a component of the inner membrane of the nuclear envelope, with the genome of glial cells of the Drosophila larval central brains.



Estimation of fluorescence lifetime of lipofuscin fluorophores contained in lipofuscin granules of retinal pigment epithelium of human cadaver eyes without signs of pathology
Abstract
The fluorescence lifetimes of lipofuscin fluorophores contained in chloroform extracts from retinal pigment epithelium (RPE) of human cadaver eyes without signs of pathology were evaluated by single photon counting. The comparison of fluorescence lifetimes of N-retinylidene-N-retinylethanolamine (A2E) and its photooxidation and photodegradation products has been carried out. It was shown that the contribution of A2E to the total fluorescence of chloroform extract from lipofuscin granules is not major. The results are important for the improvement of noninvasive diagnostic method of degenerative diseases of the retina and RPE—fundus autofluorescence (FAF).



Molecular mechanisms of synthesis of noradrenaline as an inducer of development in the adrenal glands of rats in ontogenesis
Abstract
The level of gene expression and the protein content of tyrosine hydroxylase and dopamine β-hydroxylase were determined. In the perinatal period of rats, when noradrenaline functions as a morphogenetic factor, the level of gene expression of these enzymes increased and the content of protein products of these genes was almost unchanged, indicating the difference in the regulatory mechanisms of their transcription and translation.



Inhibition of P-glycoprotein stimulates cell death under Hypoxia-mimicking conditions
Abstract
The most common drug resistance mechanism in tumor cells is expression on their surface of the energy-dependent pump like P-glycoprotein (P-gp) that expels chemotherapeutic agents from the interior. An imitation of the hypoxic condition by the iron chelator deferoxamine caused Hypoxia-inducible factor 1-alpha (HIF-1α) stabilization and inhibition of doxorubicin-induced apoptosis in colon cancer НСТ116 cells. P-gp blocker verapamil suppressed doxorubicin accumulation leading to cell death induction. Considering these results, P-gp may be used as a potential target to stimulate chemotherapeutic drugs activity that will contribute to more efficient tumor cells elimination.



Analysis of fatty acid composition revealed differences in the diets of tadpoles of two amphibian species
Abstract
The study of the composition of fatty acid markers of tadpoles of cohabiting amphibian species for the first time revealed differences in their diets: the moor frog Rana arvalis prefers bacteria not associated with plant detritus, whereas the diet of the common spadefoot Pelobates fuscus is based on cyanobacteria, green algae, diatoms, and possibly higher plants. Major differences in the fatty acid composition are determined by the difference in the percentage of eicosapentaenoic and myristic acids.



Imbalance of morphofunctional responses of Jurkat T lymphoblasts at short-term culturing with relief zinc- or copper-containing calcium phosphate coating on titanium
Abstract
Morphofunctional response of Jurkat T cells that were cultured for 24 h on substrates prepared from commercially pure titanium with relief microarc bilateral calcium phosphate coating containing copper or zinc was studied. Changes in the concentration of essential trace elements contained in this coating can cause significant imbalance of molecular processes of differentiation, secretion, apoptosis, and necrosis and reduce tumor cell survival.



Modification of the catalytic subunit of plasma fibrin-stabilizing factor under induced oxidation
Abstract
For the first time, by using mass-spectrometry method, the oxidation-mediated modification of the catalytic FXIII-A subunit of plasma fibrin-stabilizing factor, pFXIII, has been studied. The oxidative sites were identified to belong to all structural elements of the catalytic subunit: the β-sandwich (Tyr104, Tyr117, and Cys153), the catalytic core domain (Met160, Trp165, Met266, Cys328, Asp352, Pro387, Arg409, Cys410, Tyr442, Met475, Met476, Tyr482, and Met500), the β-barrel 1 (Met596), and the β-barrel 2 (Met647, Pro676, Trp692, Cys696, and Met710), which correspond to 3.9%, 1.11%, 0.7%, and 3.2%, respectively, of oxidative modifications as compared to the detectable amounts of amino acid residues in each of the structural domains. Lack of information on some parts of the molecule may be associated with the spatial unavailability of residues, complicating analysis of the molecule. The absence of oxidative sites localized within crucial areas of the structural domains may be brought about by both the spatial inaccessibility of the oxidant to amino acid residues in the zymogen and the screening effect of the regulatory FXIII-B subunit.



Silica-containing inclusions in the cytoplasm of diatom Synedra acus
Abstract
Cells of the araphid pennate diatom Synedra acus subsp. radians contain large inclusion (1–15 μm) storing silica, as revealed by transmission electron microscopy and EDX-analysis. The size of the inclusions increases with the time of cultivation of diatoms. Approximate concentration of SiO2 in the inclusions is 1–4%. It is assumed that silica in the inclusions is present as gelatinized silica-gel. These results explain the possible mechanism of transport of silicic acid into the cell against the concentration gradient.



The bxdPRE/TRE element terminates passing through transcription
Abstract
The regulation of PRE/TRE activity is required for appropriate tissue and stage-specific gene expression. However, the molecular principles of PRE/TRE activity control remain unknown. Here we show that PRE/TRE element from Ubx regulatory region efficiently terminates passing through transcription.



N-methyl serotonin analogues from the Bufo bufo toad venom interact efficiently with the α7 nicotinic acetylcholine receptors
Abstract
Two low-molecular-weight compounds were isolated from the parotid gland secret of the toad Bufo bufo, which by absorption spectra and HPLC-MS/MS chromatography data correspond to di- and trimethyl derivatives of serotonin (5-hydorxytryptamine): bufotenine (confirmed by counter synthesis) and bufotenidine (5-HTQ). In experiments on competitive radioligand binding, these compounds showed a higher affinity and selectivity for neuronal α7 nicotinic acetylcholine receptors compared with the muscular cholinergic receptors. The most efficient compound in terms of binding value was bufotenine, the efficiency of 5-HTQ was an order of magnitude lower, and the minimal activity was exhibited by serotonin.



Seasonal changes in proteolytic activity of calpains in striated muscles of long-tailed ground squirrel Spermophilus undulatus
Abstract
Seasonal changes in proteolytic activity and content of calpains in striated muscles of the longtailed ground squirrel Spermophilus undulatus were studied by casein zymography and Western blotting analysis. The results testify to hyperactivation of calpain proteases in the skeletal muscles of awakened animals during the “winter” activity. The observed changes are discussed in the context of adaptation of skeletal muscles of long-tailed ground squirrels to hibernation.



Bioluminescent assay for toxicological assessment of nanomaterials
Abstract
A new method for assessing biotoxicity of nanomaterials, based on the use of soluble bioluminescent coupled enzyme system NAD(P)⋅H:FMN oxidoreductase and luciferase, is proposed. The results of this study indicate a significant adverse biological effect exerted by nanoparticles at the molecular level. It was found that the most toxic nanoparticles the nanoparticles are based on copper and copper oxide, as well as single-walled carbon nanotubes and multi-walled carbon nanofibers, which are referred to hazard class II.



Detection of autoantibodies to potentially amyloidogenic protein, gamma-synuclein, in the serum of patients with amyotrophic lateral sclerosis and cerebral circulatory disorders
Abstract
In this study, we analyzed serum for the presence of antibodies to gamma-synuclein in patients with amyotrophic lateral sclerosis (ALS) compared to the control group of patients with other neurological diseases and healthy control donors. As a result, antibodies against gamma-synuclein are not an ALS-specific feature and have been identified in patients with ALS as well as in the control group patients. Patients with the impaired cerebral circulation showed increased incidence of autoantibodies to gamma-synuclein, yet the difference lacks statistical representativeness due to limited sample size.



PRE/TRE elements act as transcription activators in Drosophila S2 Cells
Abstract
The Drosophila PRE/TRE elements are DNA targets for Polycomb and Trithorax group proteins, which control repression and activation of gene transcription, respectively. In this study, we show that, in transiently transfected Drosophila S2 cells, bxdPRE activates transcription driven from different promoters. Using CG32795 gene promoter, we demonstrate that other PRE/TRE elements—Fab7, en, eve, and CycB—also act as transcription activators in this model system.



Long-term hyperthermia impairs activity of both photosystems
Abstract
This is the first study to perform a simultaneous analysis of the activity of photosystem 1 and photosystem 2 after long-term exposure to elevated temperature. It was found that the quantum yield of photochemical reactions decreases in both photosystems. It is shown that, in photosystem 2, the regulated nonphotochemical quenching decreases whereas the unregulated non-photochemical quenching increases. In photosystem 1, limitation on both acceptor and donor sides increases.



Sigma-1 receptor antagonist haloperidol attenuates Ca2+ responses induced by glutoxim and molixan in macrophages
Abstract
Using Fura-2AM microfluorimetry, we have shown for the first time that sigma-1 receptor antagonist, antipsychotic haloperidol, significantly inhibits glutoxim- and molixan-induced Ca2+-response in peritoneal macrophages. These results indicate possible involvement of sigma-1 receptors in the signal cascade induced by glutoxim or molixan and leading to intracellular Ca2+ concentration increase in macrophages.



Mathematical model of polymerase chain reaction with temperature-dependent parameters
Abstract
The course of the real-time polymerase chain reaction (PCR) is determined by the temperature dependence of the kinetics of the component reactions, particularly the DNA strand hybridization. To investigate the effect of thermal processes on the reaction behavior, a mathematical model in which the variable rate constant of dissociation of “primer–single strand” complexes depends on temperature was proposed. The reaction medium temperature, which depends on time, was also introduced into the model. The proposed model of real-time PCR makes it possible to analyze different aspects of the reaction, which are important for the development of instruments and reagents for PCR.



Characterization of new modular nanotransporters with albumin-binding domain
Abstract
The albumin-binding domain (ABD) with a site for its cleavage by tumor proteases was inserted in the structure of modular nanotransporters (MNTs), chimeric proteins for the delivery of anticancer drugs into the nuclei of cancer cells. The effectiveness of this cleavage was tested in both variants of created construct: “pure” ABD–MNT and the complex with albumin. The introduction of the ABD module into MNTs had no effect on the binding of MNT with receptors on the surface of the target cancer cells and on the preferential accumulation of MNTs in the nuclei of these cells. The use of thermophoresis allowed us to determine the equilibrium dissociation constants of the ABD-MNT complex with bovine and human serum albumins.


